Preparative regimen for engraftment, growth and differentiation of non-hematopoeitic cells in vivo after transplantation

Abstract

Disclosed are methods of obtaining an expanded population of mammalian ex vivo cells for treating a mammalian subject by (a) administering to a subject an effective amount of an agent that confers a growth disadvantage to at least a subset of endogenous cells at the site of engraftment; (b) administering to the subject an effective amount of a mitogenic stimulus for the ex vivo cells; and (c) administering the ex vivo cells to the subject, wherein the ex vivo cells engraft at the site and proliferate to a greater extent than the subset of endogenous cells, to repopulate at least a portion of the engraftment site with the ex vivo cells. The repopulated cells can be harvested or left at the engraftment site. Methods of treating brain injury in a subject by engrafting ex vivo cells at the site of injury are also described.

1 . A method of treating a mammalian subject with an organ having a reduced function by grafting mammalian ex vivo cells which supplement the function and by promoting proliferation of the engrafted cells at the site of engraftment, the method comprising the steps of:
 (a) administering to the subject an effective amount of an agent that confers a growth disadvantage to at least a subset of endogenous cells at the site of engraftment;   (b) administering to the subject an effective amount of a mitogenic stimulus for the ex vivo cells; and   (c) administering the ex vivo cells to the subject, wherein the ex vivo cells engraft at the site and proliferate to a greater extent than the subset of endogenous cells, to repopulate at least a portion of the engraftment site with the ex vivo cells, wherein the repopulated ex vivo cells supplement or provide the function.   
     
     
         2 . The method of  claim 1 , wherein the organ with the reduced function is the intestine, liver, lung, kidney, pancreas, eye, testes, ovary, brain, spinal cord, or skin. 
     
     
         3 . (canceled) 
     
     
         4 . (canceled) 
     
     
         5 . The method of  claim 1 , wherein the ex vivo cells are selected from the group consisting of adult somatic cells, adult progenitor cells, adult stem cells, embryonic progenitor cells, fetal cells, and embryonic stem cells from the same species as the subject or from a different mammalian species than the subject. 
     
     
         6 . The method of  claim 1 , wherein the ex vivo cells are heterologous or autologous to the subject, are cultured, expanded or modified prior to their administration to the subject, or are harvested and sorted from a heterogeneous cell population prior to their administration to the subject. 
     
     
         7 . (canceled) 
     
     
         8 . (canceled) 
     
     
         9 . (canceled) 
     
     
         10 . The method of  claim 1 , wherein the agent that confers a growth disadvantage to at least a subset of endogenous cells is radiation, x-ray, gamma ray, or is selected from the group consisting of cytotoxic chemicals, ultrasound, heat, biological agents, misfolded proteins, and proteins or nucleic acids that suppress cell division. 
     
     
         11 . (canceled) 
     
     
         12 . (canceled) 
     
     
         13 . (canceled) 
     
     
         14 . (canceled) 
     
     
         15 . The method of  claim 1 , wherein the administered ex vivo cells have been genetically modified or comprise a heterologous gene that increases their intrinsic proliferation capacity or survival. 
     
     
         16 . The method of  claim 1 , wherein the mitogenic stimulus comprises biological agents, antibodies and peptide factors that increase the proliferation capacity of the donor cells or a procedure that deliberately injures the engraftment site to enable or to promote entry and integration of the engrafted ex vivo cells into the host parenchyma and/or to provide a compensatory growth signal for the ex vivo cells. 
     
     
         17 . (canceled) 
     
     
         18 . (canceled) 
     
     
         19 . (canceled) 
     
     
         20 . (canceled) 
     
     
         21 . The method of  claim 1 , wherein the ex vivo cells are heterologous to the engraftment site. 
     
     
         22 . The method of  claim 1 , wherein the engraftment site is the organ having the reduced function, or the engraftment site is different from the organ having the reduced function. 
     
     
         23 . (canceled) 
     
     
         24 . (canceled) 
     
     
         25 . The method of  claim 1 , wherein the ex vivo cells express a heterologous nucleic acid for gene therapy. 
     
     
         26 . (canceled) 
     
     
         27 . (canceled) 
     
     
         28 . (canceled) 
     
     
         29 . (canceled) 
     
     
         30 . The method of  claim 1 , wherein the ex vivo cell expresses a protein not expressed by the host tissue at the engraftment site and the presence of the protein is used to selectively detect the ex vivo cells as opposed to the host cells at the engraftment site. 
     
     
         31 . (canceled) 
     
     
         32 . A method of obtaining an expanded population of ex vivo cells, comprising the steps of:
 (a) administering to a non-human mammalian subject an effective amount of an agent that confers a growth disadvantage to at least a subset of endogenous cells at a site of engraftment;   (b) administering to the non-human mammalian subject an effective amount of a mitogenic stimulus for the ex vivo cells; and   (c) administering the ex vivo cells to the subject, wherein the ex vivo cells engraft at the site and proliferate to a greater extent than the subset of endogenous cells, thereby repopulating at least a portion of the engraftment site with the ex vivo cells, and (d) harvesting the repopulated cells from the engraftment site;   thereby obtaining the expanded ex vivo cell population.   
     
     
         33 . The method of  claim 32 , wherein the ex vivo cells are human. 
     
     
         34 . The method of  claim 32 , wherein the ex vivo cells are selected from the group consisting of adult somatic cells, adult progenitor cells, adult stem cells, embryonic progenitor cells, fetal cells, xenogenic cells, and embryonic stem cells. 
     
     
         35 . The method of  claim 32 , wherein the ex vivo cells heterologous to the non-human subject. 
     
     
         36 . The method of  claim 32 , wherein the agent that confers a growth disadvantage to at least a subset of endogenous cells of the organ is radiation, x-ray, gamma ray, or is selected from the group consisting of cytotoxic chemicals, ultrasound, heat, biological agents, misfolded proteins, and proteins or nucleic acids that suppress cell division. 
     
     
         37 . (canceled) 
     
     
         38 . (canceled) 
     
     
         39 . (canceled) 
     
     
         40 . The method of  claim 32 , wherein the ex vivo cell is derived from brain, intestinal, or liver tissue. 
     
     
         41 . (canceled) 
     
     
         42 . (canceled) 
     
     
         43 . (canceled) 
     
     
         44 . A method of obtaining an expanded population of ex vivo cells, comprising the steps of:
 (a) administering to an irradiated organ ex vivo or in culture an effective amount of a mitogenic stimuli for the ex vivo cells; wherein the irradiation confers a growth disadvantage to at least a subset of endogenous cells at a site where the ex vivo cells engraft; and   (b) administering the ex vivo cells to the irradiated organ, wherein the ex vivo cells engraft at the site and proliferate to a greater extent than the subset of endogenous cells, thereby repopulating at least a portion of the engraftment site with the ex vivo cells, and   (c) harvesting the repopulated cells from the engraftment site;   
       thereby obtaining the expanded ex vivo cell population. 
     
     
         45 - 52 . (canceled)